A novel small-molecule inhibitor of the chromosome segregation process in yeast
- 77 Downloads
The centromeres of budding yeasts contain specific and essential DNA sequences, in contrast to the regional heterochromatic centromeres found in higher organisms. Small molecules that perturb centromere function in budding yeast could be valuable candidates for identifying yeast-specific growth inhibitors. A combination of two in vivo assays, one based on transcription blockade of a reporter gene by the centromeric DNA-protein complex, the other on a test for mitotic minichromosome stability, was used to identify small molecules that affect the process of chromosome segregation. One compound, here named incentrom A, leads to a minichromosome loss phenotype, and is cytotoxic to the budding yeast Saccharomyces cerevisiae. Furthermore, whereas cells carrying a conditional mutation in a gene for an essential kinetochore protein, skp1-4 , are hypersensitive to incentrom A, cells that overexpress the SKP1 gene are resistant to the compound. Incentrom A also results in mitotic loss of a centromere-bearing plasmid, and inhibits the growth of the pathogenic yeast Candida glabrata. Incentrom A will therefore be a useful tool for studying the molecular basis of yeast chromosome segregation and could form the basis for the development of novel antifungal drugs.
KeywordsCentromere Chromosome segregation Incentrom Small-molecule inhibitor Antifungal drug
I am grateful to John Carbon for critical reading of the manuscript. I thank H. Roe and E. George for technical assistance, past and present members of the Carbon and Clarke laboratories for many discussions, and P. Hieter, C. Dougherty, A. Murray, D. Gebow and T. Stoyan for providing yeast strains and/or plasmids. I thank D. Gebow for sharing useful ideas and S. Shin for obtaining the mass spectrum of incentrom A. A portion of this work was carried out in John Carbon’s laboratory at the University of California, Santa Barbara, and was supported by NIH Grant CA-11034. This work was also supported by the Brain Korea 21 Project, the KISTEP NRL Program (Grant No. 2000-N-NL-01-C-210), and the KIST Functional Proteomics Center.
- Andreeva NI, Mashkovsky MD, Golovina SM, Gorkin VZ (1992) Novel antidepressant drug tetrindole (2, 3, 3a, 4, 5, 6-hexahydro-8-cyclohexyl-1H-pyrazino [3, 2, 1-j, k] carbazole. 1. Pharmacological study of the influence on central nervous system. Chem Pharm J 26:17–23Google Scholar
- Centers for Disease Control and Prevention (2001) Candidiasis information. Division of bacterial and mycotic diseasesGoogle Scholar
- Gardner RD, Poddar A, Yellman C, Tavormina PA, Monteagudo MC, Burke DJ (2001) The spindle checkpoint of the yeast Saccharomyces cerevisiae requires kinetochore function and maps to the CBF3 domain. Genetics 157:1493–1502Google Scholar
- Hieter P, Mann C, Snyder M, Davis RW (1985) Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss. Cell 40:381–392Google Scholar
- Ng R, Cumberledge S, Carbon J (1986) Structure and function of centromeres. In: Hicks J (ed) Yeast cell biology. Alan R. Liss, New York, pp 225–239Google Scholar
- Pfaller MA, Messer SA, Hollis RJ, Jones RN, Doern GV, Brandt ME, Hajjeh RA (1999) Trends in species distribution and susceptibility to fluconazole among blood stream isolates of Candida species in the United States. Diagn Microbiol Infect Dis 33:217–222Google Scholar
- Schreiber SL (2003) The small-molecule approach to biology. Chem Eng News 81:51–61Google Scholar